title: A fluorescence-based high-throughput screening assay to identify HIV-1 inhibitors.
abstract: Highly active antiretroviral therapy (HAART) dramatically increases the long-term survival rates of human immunodeficiency virus type 1 (HIV-1) infected patients. Yet, poor adherence to therapy, adverse effects and the occurrence of resistant viruses can compromise the efficacy of HAART regiments. Therefore, there remains a clear unmet medical need for novel drugs and treatment options. In this chapter, we describe an HIV-1 antiviral high-throughput screening assay based on an HIV-1 permissive T lymphoblastoid MT4 cell line, stably transfected with a construct carrying an HIV-1 long terminal repeat promoter driving the expression of a reporter gene (enhanced green fluorescent protein). This assay runs in a 384-well format and enables the identification of HIV-1 inhibitors during a high-throughput screening campaign. In parallel, a cytotoxicity assay is performed to evaluate the compound-related in vitro toxicity.
authors.affiliation: Janssen Infectious Diseases-Diagnostics BVBA, Johnson & Johnson Corporation, Beerse, Belgium.
authors.forename: Peggy
authors.initials: P
authors.lastname: Geluykens
authors.forename: Koen
authors.initials: K
authors.lastname: Van Acker
authors.forename: Johan
authors.initials: J
authors.lastname: Vingerhoets
authors.forename: Christel
authors.initials: C
authors.lastname: Van den Eynde
authors.forename: Marnix
authors.initials: M
authors.lastname: Van Loock
authors.forename: Géry
authors.initials: G
authors.lastname: Dams
mesh_terms: D019380:Anti-HIV Agents / Q000494:pharmacology*; D015870:Gene Expression; D017930:Genes, Reporter; D015497:HIV-1 / Q000187:drug effects*; D057166:High-Throughput Screening Assays / Q000379:methods; D006801:Humans; D008856:Microscopy, Fluorescence; D014779:Virus Replication / Q000187:drug effects
chemical_list: D019380:Anti-HIV Agents